Preparation method and applications of rapeseed cake-sourced metal chelating peptide and peptide metal chelate

Abstract

The invention belongs to the technical field of deep processing of rapeseed cake byproducts, and particularly relates to a preparation method and applications of rapeseed cake-sourced metal chelating peptide and peptide metal chelate. The preparation method comprises the steps: soaking and cleaning rapeseed cakes with ethanol and water in sequence, and performing solid-liquid separation to reserve a filter cake, thus obtaining a crude rapeseed protein product; adding water into the crude rapeseed protein product, regulating the pH value, then adding protease for hydrolysis reaction, then placing reaction liquid into boiling water for cooking and carrying out enzyme deactivation, to obtain hydrolysate; or, adding the protease for hydrolysis reaction, then placing reaction liquid into an autoclave for high-pressure treatment, to obtain the hydrolysate with higher hydrolysis degree; and centrifuging the hydrolysate, and drying supernate to obtain crude metal chelating peptide powder. The prepared metal chelating peptide prepared by the preparation method disclosed by the invention can be directly used as a natural antioxidant for retarding aging. In addition, the metal chelating peptide can also be used for preparing an excellent parent of the peptide metal chelate, and can be chelated with different microelements to prepare a micro trace chelating agent for supplementing nutrient elements which are lack by the human body.

Description

technical field

[0001] The invention belongs to the technical field of deep processing of rapeseed cake by-products, and in particular relates to a preparation method and application of a rapeseed cake source metal chelating peptide and a peptide metal chelate. Background technique

[0002] Metal chelating peptides refer to bioactive peptides that can chelate metal ions. Metal ions such as copper, zinc, and iron are widely present in the organs and tissues of the body. When there are excessive metal ions in the body, it is easy to lead to the generation of reactive oxygen species (Reactive oxygen species, ROS), and then damage the biomacromolecules in the body, resulting in some degenerative diseases, such as cardiovascular and cerebrovascular diseases, gastrointestinal inflammation and cancer Wait. Metal chelating peptide itself can chelate free metal ions in the body, and play a role in inhibiting or reducing the oxidative damage caused by active oxygen in the body. In ...

Examples

Embodiment 1

[0026] The rapeseed cake was pulverized by using a high-speed pulverizer, 500 g of raw material was taken, 3 L of 75% (volume percentage) ethanol aqueous solution was added, stirred and soaked for 24 hours, and the liquid was changed every 6 hours during this period. After filtering, the precipitate was washed with 3L of water and then filtered, and after drying, a light brown crude rapeseed protein was obtained. The average yield of crude protein was 21.46%.

Embodiment 2

[0028] Take 4g of rapeseed crude protein, add 100ml of deionized water, adjust the pH to 8.0 with 1mol / L NaOH solution, add 0.08g of Alcalase enzyme, and carry out the enzymatic hydrolysis reaction of the reaction mixture at 55°C, during which 1mol / L L of NaOH solution was used to adjust the pH to maintain the reaction system at a pH of 8.0. When the reaction time reaches 4 hours, put the reaction solution in a boiling water bath to inactivate the enzyme, then put the protein hydrolyzate in a pressure cooker, and treat it under high pressure at 121° C. for 10 minutes to obtain a hydrolyzate.

[0029] The obtained high-pressure treatment hydrolyzate is subjected to freeze-drying treatment to obtain crude metal chelating peptide powder. The degree of hydrolysis of the hydrolyzate was 22.31% when detected by TNBS method for 4 hours. When the peptide concentration was 5 mg / ml, its Zn was detected by EDTA titration 2+ The average chelation rate is 84.36%.

Embodiment 3

[0031] Take 4g of rapeseed crude protein, add 100ml of deionized water, adjust the pH to 7.0 with 1mol / L NaOH solution, add 0.08g of papain, and carry out the enzymatic hydrolysis reaction of the reaction mixture at 60°C, during which 1mol / L L of NaOH solution was used to adjust the pH to maintain the reaction system at a pH of 7.0. When the reaction time reaches 1 hour, put the hydrolyzate in boiling water and inactivate the enzyme for 10 minutes.

[0032] The obtained hydrolyzate is freeze-dried to obtain crude metal chelating peptide powder. The degree of hydrolysis of the hydrolyzate was 20.82% when detected by TNBS method for 1 hour. When the peptide concentration was 5 mg / ml, its Zn was detected by EDTA titration 2+ The average chelation rate is 29.79%.

technical field

[0001] The invention belongs to the technical field of deep processing of rapeseed cake by-products, and in particular relates to a preparation method and application of a rapeseed cake source metal chelating peptide and a peptide metal chelate. Background technique

[0002] Metal chelating peptides refer to bioactive peptides that can chelate metal ions. Metal ions such as copper, zinc, and iron are widely present in the organs and tissues of the body. When there are excessive metal ions in the body, it is easy to lead to the generation of reactive oxygen species (Reactive oxygen species, ROS), and then damage the biomacromolecules in the body, resulting in some degenerative diseases, such as cardiovascular and cerebrovascular diseases, gastrointestinal inflammation and cancer Wait. Metal chelating peptide itself can chelate free metal ions in the body, and play a role in inhibiting or reducing the oxidative damage caused by active oxygen in the body. In ...

Examples

Embodiment 1

[0026] The rapeseed cake was pulverized by using a high-speed pulverizer, 500 g of raw material was taken, 3 L of 75% (volume percentage) ethanol aqueous solution was added, stirred and soaked for 24 hours, and the liquid was changed every 6 hours during this period. After filtering, the precipitate was washed with 3L of water and then filtered, and after drying, a light brown crude rapeseed protein was obtained. The average yield of crude protein was 21.46%.

Embodiment 2

[0028] Take 4g of rapeseed crude protein, add 100ml of deionized water, adjust the pH to 8.0 with 1mol / L NaOH solution, add 0.08g of Alcalase enzyme, and carry out the enzymatic hydrolysis reaction of the reaction mixture at 55°C, during which 1mol / L L of NaOH solution was used to adjust the pH to maintain the reaction system at a pH of 8.0. When the reaction time reaches 4 hours, put the reaction solution in a boiling water bath to inactivate the enzyme, then put the protein hydrolyzate in a pressure cooker, and treat it under high pressure at 121° C. for 10 minutes to obtain a hydrolyzate.

[0029] The obtained high-pressure treatment hydrolyzate is subjected to freeze-drying treatment to obtain crude metal chelating peptide powder. The degree of hydrolysis of the hydrolyzate was 22.31% when detected by TNBS method for 4 hours. When the peptide concentration was 5 mg / ml, its Zn was detected by EDTA titration 2+ The average chelation rate is 84.36%.

Embodiment 3

[0031] Take 4g of rapeseed crude protein, add 100ml of deionized water, adjust the pH to 7.0 with 1mol / L NaOH solution, add 0.08g of papain, and carry out the enzymatic hydrolysis reaction of the reaction mixture at 60°C, during which 1mol / L L of NaOH solution was used to adjust the pH to maintain the reaction system at a pH of 7.0. When the reaction time reaches 1 hour, put the hydrolyzate in boiling water and inactivate the enzyme for 10 minutes.

[0032] The obtained hydrolyzate is freeze-dried to obtain crude metal chelating peptide powder. The degree of hydrolysis of the hydrolyzate was 20.82% when detected by TNBS method for 1 hour. When the peptide concentration was 5 mg / ml, its Zn was detected by EDTA titration 2+ The average chelation rate is 29.79%.

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